基于PCA和PLS-DA算法的不同生长阶段贯叶金丝桃药材HPLC指纹图谱研究

目的：建立不同生长阶段贯叶金丝桃HPLC指纹图谱，并结合相似度计算和化学模式识别技术对其进行质量评价。方法：采用Waters Sunfire ODS（250 mm×4.6 mm，5 μm）色谱柱；乙腈-0.05%磷酸水溶液为流动相，梯度洗脱，流速1.0 mL·min^-1，检测波长254 nm，建立18批贯叶金丝桃的指纹图谱。采用相似度分析、主成分分析（PCA）及偏最小二乘法-判别分析（PLS-DA）对其总体质量进行分析评价。结果：建立了贯叶金丝桃指纹图谱且共标定16个共有峰，经过与对照品比对，指认并鉴定出其中的6个色谱峰。18批样品的相似度为0.712~0.933，PCA分析出不同生长阶段的贯叶金丝桃药材的最佳采收阶段为盛开期；PLS-DA可将不同生长阶段的样品显著分类，并筛选出导致不同生长阶段质量差异的6个差异标志物。结论：该研究建立的指纹图谱分析方法科学、准确，结合PCA和PLS-DA化学模式识别技术，可系统、全面地评价贯叶金丝桃的质量。

Objective: To establish an HPLC fingerprint of Hyperici Perforati Herba at different growth stages,and to evaluate the quality through the similarity calculation and chemical pattern recognition. Methods: The analysis was performed on a Waters Sunfire ODS(250 mm×4.6 mm,5 μm)column with mobile phase consisting of acetonitrile-0.05% phosphoric acid solution(gradient elution)at a flow rate of 1.0 mL·min^-1. The detection wavelength was set at 254 nm to establish the HPLC fingerprints of the 18 batches of Hyperici Perforati Herba. Then,the further quality assessment of the herb was carried out by similarity avaluation,principal component analysis(PCA),and discriminant analysis of partial least squares(PLS-DA). Results: HPLC fingerprint of Hyperici Perforati Herba was established and 16 common peaks were identified with 6 peaks were identified using standard references. The similarity calculation results were 0.712-0.933 and PCA showed that the optimum harvesting stage of Hyperici Perforati Herba was the blooming stage. And PLS-DA screened out 6 differential markers that led to quality differences at different growth stages. Conclusion: The fingerprint analysis method established in this study is scientific and accurate. Combining PCA and PLS-DA chemical pattern recognition technology,the quality of Hyperici Perforati Herba can be evaluated systematically and comprehensively.

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